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Figure 2 | BMC Cancer

Figure 2

From: MiR-200c and HuR in ovarian cancer

Figure 2

A: Expression analysis of miR-200c in A2780 cells transformed with the empty vector (pUSE) and in two clones overexpressing the miR-200c (200c-5 and 200c-14). B: Bar chart of TUBB3 mRNA expression obtained in the same cells. Bars and error bars refer to mean and SD of two experiments performed in triplicate. It is clearly visible that overexpression of miR-200c inhibits expression of TUBB3 gene in a significant way (double asterisks= P<0.001). The same phenomenon is evident in the western blot (C) showing the TUBB3 expression at the protein level. Actin served as a loading control. D: Schematic representation of the constructs utilized for the luciferase assays. E: Luciferase assays on A2780 cells transfected with the reporter vectors pGL3-TUBB3-UTR, pGL3-TUBB3-UTRm or the control pGL3-V. The relative luciferase units were calculated normalizing the firefly values for the renilla values. A clear inhibition is noticeable when pGL3-TUBB3-UTR is transfected in miR-200c is overexpressing cells . F-G: Line charts reporting dose–response curves obtained in A2780-pUSE, 200c-5 and 200c-14 cells treated with paclitaxel (F) and cisplatinum (G). Colonies were counted 14 d after drug treatment.

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